Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.4.21.1 (chymotrypsin)
 10,938 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Our previous studies shown that a high caloric load in the jejunum decreases biliopancreatic output. However, the factors responsible for this inhibition have not yet been fully assessed. In this study, we have compared the effect of a high caloric load of either proteins or carbohydrates on stimulated pancreatic output and investigated the mechanism of this inhibition. The small intestine of 6 healthy volunteers was intubated 10 cm below the angle of Treitz, for 3 consecutive days, to infuse test solutions. After a 90 min period of stimulation, using a protein solution (0.6 kcal/min; solution A), one of the following 2 solutions was infused on for 90 min each day, in random order. In each instance, either a protein plus carbohydrate solution (3.5 kcal/min; solution B), or a protein solution (3.5 kcal/min; solution C) was administered. On the 3rd day, first solution A then solution B were infused with either IV atropine (17 mug/kg/h) in 3 subjects, or naloxone (40 mug/kg/hl) in 3 subjects. PEG 4000 was also added to solutions B and C and (14)C-PEG to solution A, as nonabsorbable markers. Intraluminal content was aspirated 25 cm below the infusion point and 50 cm distally to prevent unabsorbed nutrients from reaching the ileum. Solution B and C caused similar decreases in lipase, chymotrypsin and bile salt outputs. Neither atropine nor naloxone inhibited the jejunal-brake induced by solution B. However, naloxone, but not atropine, decreased the carbohydrate absorption rate of solution B. In conclusion, a high caloric load of proteins or carbohydrates infused into the human jejunum decreased the bilio-pancreatic output induced by a low caloric load of proteins. This mechanism is neither cholinergic nor opioid dependent. This effect, in relation to a jejunal brake, might explain functional pancreatic insufficiency after surgical procedures such as gastro-jejunostomy.
 ...
PMID:Inhibitory effect of jejunal high caloric nutrient load on human biliopancreatic secretion. The role of atropine, naloxone and composition of nutrient solutions. 1684 72

Metaxalone (Met), a drug for treatment of pain and stiffness due to muscular injuries, was covalently linked to poly(ethylene glycols) (PEG) via a chloroacetyl chloride spacer. The average weight molecular weights used for PEG are 4000, 6000 and 10,000, respectively, and the procedure of chemical modification for PEGs was conducted by a two-step protocol: (1) synthesis of N-chloroacetyl-metaxalone; (2) synthesis of PEG(4000)-Met, PEG(6000)-Met and PEG(10000)-Met. The controlled drug release studies were performed in buffer solutions with pH values equal to 1.1, 7.4 and 10.0. The results demonstrate that, in the same condition, the rate of hydrolysis for PEG(10000)-Met is the slowest among three prodrugs, and more amount of metaxalone can be detected releasing from prodrug matrices at the presence of alpha-chymotrypsin in a buffer solution with pH 8.0. It was also found that these novel prodrugs can effectively improve the metaxalone's pharmacokinetics, and furthermore can markedly increase its half-life period.
 ...
PMID:Synthesis of poly(ethylene glycol)-metaxalone conjugates and study of its controlled release in vitro. 1706 68

The partitioning of bovine trypsin and alpha-chymotrypsin--proteases of similar physico-chemical properties--in different polyethyleneglycol/sodium citrate aqueous two-phase systems was investigated. The effect of different factors such as polyethyleneglycol molecular weight, pH, tie line length, temperature and the presence of an inorganic salt on the protein partition coefficient were analysed. Both a decrease in PEG molecular weight and an increase in pH led to a higher partition coefficient for both enzymes. Aqueous two-phase systems formed by PEG of molecular weight 3350 and citrate pH 5.2 showed the best separation capability which was enhanced in presence of sodium chloride 3%. The transfer of both proteins to the top phase was associated with negative enthalpic and entropic changes.
 ...
PMID:Partitioning features of bovine trypsin and alpha-chymotrypsin in polyethyleneglycol-sodium citrate aqueous two-phase systems. 1730 4

The biodegradability of both unsaturated (UPEA) and saturated (SPEA) poly(ester-amide)s and a series of hydrogels (UPEA-G) fabricated from UPEA and poly(ethylene glycol) diacrylate (PEG-DA) was examined as a function of PEA chemical structures in both phosphate buffered saline (PBS) and alpha-chymotrypsin solutions. Based on the weight loss data, alpha-chymotrypsin had a much more profound effect on the hydrolyses of UPEA, SPEA polymers (up to 32% weight loss on day 1 for FPBe) and UPEA-G hydrogels (up to 32% weight loss on day 31 for FPBe-G28) than a PBS buffer (less than 10% for polymers and 16% for hydrogels). The changes in elastic moduli and the interior morphology of the hydrogels in both PBS buffer and alpha-chymotrypsin solutions were also monitored for 2 months, and the hydrogels' crosslinking density (n(e)) and molecular weight between crosslinks (M(c)) before and after biodegradation were then examined as a function of biodegradation time, enzyme concentration, and different chemical structure of precursors. The differences in biodegradation rates among PEA polymer and UPEA-G hydrogels are ascribed to differences in hydrophilicity and saturated or unsaturated structure of the polymers and hydrogel precursors. Our results showed that, by changing the concentration of alpha-chymotrypsin, the type of UPEA precursors and their feed ratio, the UPEA-G hydrogels could have controllable biodegradability, which is quite desirable for a wide range of biomedical and pharmaceutical applications.
 ...
PMID:Biodegradation of unsaturated poly(ester-amide)s and their hydrogels. 1746 69

Biodegradable hydrogels (FPBe-G) were synthesized by the photopolymerization of two precursors: FPBe, a fumurate-based unsaturated poly(ester amide) (UPEA), and poly(ethylene glycol) diacrylate (PEG-DA). Depending on the feed ratio of these two precursors, the resultant FPBe-G hydrogels showed different crosslinking levels of network structure, mesh sizes (xi) and matrix morphology. When a lipophilic drug, paclitaxel, was preloaded into FPBe-G hydrogels, the two-month drug-release kinetics from FPBe-G hydrogels in both pure PBS buffer and alpha-chymotrypsin media were measured. The paclitaxel-preloaded FPBe-G hydrogels in a alpha-chymotrypsin solution had significantly faster drug release rate than the corresponding hydrogels in a pure PBS buffer due to an enzyme catalyzed biodegradation of FPBe-G hydrogels. Sustained paclitaxel releases over a two-month period without initial burst release were also achieved by using hydrogels having certain feed ratios of hydrogel precursors. These paclitaxel release data correlated well with the molecular mesh size (xi), molecular weight between cross-links (M(c)) and matrix morphological structure of FPBe-G hydrogels.
 ...
PMID:Controlled release of paclitaxel from biodegradable unsaturated poly(ester amide)s/poly(ethylene glycol) diacrylate hydrogels. 1755 Jun 54

SPCI, a Kunitz-type chymotrypsin inhibitor, is a 180-amino-acid polypeptide isolated from Schizolobium parahyba seeds. This inhibitor has been characterized as a highly stable protein over a broad pH and temperature range. SPCI was crystallized using a solution containing 0.1 M sodium acetate trihydrate buffer pH 4.6, 33%(v/v) PEG 2000 and 0.2 M ammonium sulfate. Data were collected to 1.80 A resolution from a single crystal of SPCI under cryogenic conditions. The protein crystallized in space group P2(1)2(1)2, with unit-cell parameters a = 40.01, b = 71.58, c = 108.68 A and an R(merge) of 0.052. The structure of SPCI has been solved by molecular replacement using the known structure of the Kunitz-type trypsin inhibitor from Delonix regia (PDB code 1r8n) as the search model.
 ...
PMID:Crystallization and preliminary crystallographic studies of Schizolobium parahyba chymotrypsin inhibitor (SPCI) at 1.8 A resolution. 1800 42

The reverse staining, with imidazole-SDS-zinc, of PEG-linked proteins separated by SDS-PAGE was studied. Using model conjugates (interferon-alpha 2b (IFN-alpha2b) reacted with either a branched-chain (40,000) PEG (PEG2,40) or a linear monomethoxy PEG polymer (Mr of 12,000) and chromatographically purified monoPEG2,40-IFN-alpha2b), conventional small-format analytical gels (<1 mm thick) showed typical detection patterns (i.e., transparent, colorless bands clearly discernible against a zinc imidazolate-generated white gel background), in less than 20 min. Nonreacted (free) PEG was almost undetected, as expected. The reverse-stained PEGylated IFN-alpha2b patterns were qualitatively indistinguishable from those of parallel gels stained with iodine (I2). The LOD was estimated in the low nanogram range (e.g., at about 7 ng for mono- or bi-PEG2,40 IFN-alpha2b per lane on gradient (4-17%) gels). Also, this stain allowed the visualization of Coomassie blue-undetected PEG-IFN bands, and could be restained with I2. PEGylated species of lysozyme, a low-molecular-weight peptide, ovalbumin, and chymotrypsin were used to demonstrate the generality of this stain. We also show (i) how to counteract the adverse effect of some parameters (e.g., gel thickness above 1 mm, long gel length, low (e.g., 4-6%) acrylamide concentration) on the reverse staining process and (ii) that the properties of the reverse-stained PEGylated proteins remain unchanged, as judged by analyzing both the ion exchange chromatography-based positional isomer separation profile and enzyme-linked immunosorbent response of PEG-IFN recovered from gels. Consequently, this technique may be useful for the rapid analysis or the small-scale preparation of PEGylated proteins.
 ...
PMID:Detection of PEGylated proteins in polyacrylamide gels by reverse staining with zinc and imidazole salts. 1844 61

The haemolytic power of isolated nematocysts from the scyphozoan Pelagia noctiluca was studied with attention to the effect of osmotic protectants as carbohydrates at different MW, cations as Mg2+, Ca2+, Ba2+,Cu2+, K+; proteases as collagenase, trypsin, alpha-chymotrypsin, papain; and antioxidants. Crude venom was at first obtained by sonication of holotrichous-isorhiza nematocysts previously isolated from oral arms of P. noctiluca and then haemolytically tested upon human erythrocytes. Osmotic protectants were effective in inhibiting the haemolytic power depending on their molecular weight so that total inhibition of crude venom-induced haemolysis was observed after PEG treatment (polyethyleneglycol 6000Da). Amongst divalent cations only Ba2+ and Cu2+ significantly inhibited the haemolytic power of crude venom. Proteases seem not to alter the haemolytic activity while antioxidant compounds only slightly reduced the haemolytic power. Such findings may suggest a pore-forming mechanism for P. noctiluca crude venom rather than an oxidative damage to the cell membrane.
 ...
PMID:Effect of various factors on Pelagia noctiluca (Cnidaria, Scyphozoa) crude venom-induced haemolysis. 1861 52

Protein stability remains one of the main factors limiting the realization of the full potential of protein therapeutics. Poly(ethylene glycol) (PEG) conjugation to proteins has evolved into an important tool to overcome instability issues associated with proteins. The observed increase in thermodynamic stability of several proteins upon PEGylation has been hypothesized to arise from reduced protein structural dynamics, although experimental evidence for this hypothesis is currently missing. To test this hypothesis, the model protein alpha-chymotrypsin (alpha-CT) was covalently modified with PEGs with molecular weights (M(W)) of 700, 2,000 and 5,000 and the degree of modification was systematically varied. The procedure did not cause significant tertiary structure changes. Thermodynamic unfolding experiments revealed that PEGylation increased the thermal transition temperature (T(m)) of alpha-CT by up to 6 degrees C and the free energy of unfolding [DeltaG(U) (25 degrees C)] by up to 5 kcal/mol. The increase in stability was found to be independent of the PEG M(W) and it leveled off after an average of four PEG molecules were bound to alpha-CT. Fourier-transformed infrared (FTIR) H/D exchange experiments were conducted to characterize the conformational dynamics of the PEG-conjugates. It was found that the magnitude of thermodynamic stabilization correlates with a reduction in protein structural dynamics and was independent of the PEG M(W). Thus, the initial hypothesis proved positive. Similar to the thermodynamic stabilization of proteins by covalent modification with glycans, PEG thermodynamically stabilizes alpha-CT by reducing protein structural dynamics. These results provide guidance for the future development of stable protein formulations.
 ...
PMID:Stabilization of alpha-chymotrypsin upon PEGylation correlates with reduced structural dynamics. 1878 98

PEGylation of peptide drugs prolongs their circulating lifetimes in plasma. However, PEGylation can produce a decrease in the in vitro bioactivity. Longer poly(ethylene glycol) (PEG) chains are favourable for circulating lifetimes but unfavourable for in vitro bioactivities. In order to circumvent the conflicting effects of PEG length, a hydrophobic peptide, using an antimicrobial peptide as a model, was PEGylated with short PEG chains. The PEGylated peptides self-assembled in aqueous solution into micelles with PEG shell and peptide core. In these micelles, the core peptides were protected by the shell, thus reducing proteolytic degradation. Meanwhile, most of the in vitro antimicrobial activities still remained due to the short PEG chain attached. The stabilities of the PEGylated peptides were much higher than that of the unPEGylated peptides in the presence of chymotrypsin and serum. The antimicrobial activities of the PEGylated peptides in the presence of serum, an ex vivo assay, were much higher than that of the unPEGylated peptide.
 ...
PMID:Modification of antimicrobial peptide with low molar mass poly(ethylene glycol). 1884 67


<< Previous 1 2 3 4 5 6 7 Next >>